Reprogramming of Sertoli cells to fetal-like Leydig cells by Wt1 ablation

Significance Genetic control of the differentiation between Sertoli cells and granulosa cells has been reported previously. However, the relationship between Sertoli cells and Leydig cells in the testis has not yet been definitively determined. In the present study, we demonstrate for the first time, to our knowledge, that these two cell types can be mutually reprogrammed and that Wilms’ Tumor Gene 1 (Wt1) plays a critical role in this process. This study provides a novel concept for cell fate determination in testis development that will improve our understanding of the regulatory mechanisms of gonad development. Sertoli and Leydig cells, the two major somatic cell types in the testis, have different morphologies and functions. Both are essential for gonad development and spermatogenesis. However, whether these cells are derived from the same progenitor cells and the mechanism regulating the differentiation between these two cell types during gonad development remains unclear. A previous study showed that overactivation of Ctnnb1 (cadherin-associated protein, beta 1) in Sertoli cells resulted in Sertoli cell tumors. Surprisingly, in the present study, we found that simultaneous deletion of Wilms’ Tumor Gene 1 (Wt1) and overactivation of Ctnnb1 in Sertoli cells led to Leydig cell-like tumor development. Lineage tracing experiments revealed that the Leydig-like tumor cells were derived from Sertoli cells. Further studies confirmed that Wt1 is required for the maintenance of the Sertoli cell lineage and that deletion of Wt1 resulted in the reprogramming of Sertoli cells to Leydig cells. Consistent with this interpretation, overexpression of Wt1 in Leydig cells led to the up-regulation of Sertoli cell-specific gene expression and the down-regulation of steroidogenic gene expression. These results demonstrate that the distinction between Sertoli cells and Leydig cells is regulated by Wt1, implying that these two cell types most likely originate from the same progenitor cells. This study thus provides a novel concept for somatic cell fate determination in testis development that may also represent an etiology of male infertility in human patients.

[1]  W. Pu,et al.  WT1 maintains adrenal-gonadal primordium identity and marks a population of AGP-like progenitors within the adrenal gland. , 2013, Developmental cell.

[2]  T. Jiang,et al.  The Wilms Tumor Gene, Wt1, Is Critical for Mouse Spermatogenesis via Regulation of Sertoli Cell Polarity and Is Associated with Non-Obstructive Azoospermia in Humans , 2013, PLoS genetics.

[3]  Satoru Takahashi,et al.  Two distinct origins for Leydig cell progenitors in the fetal testis. , 2011, Developmental biology.

[4]  H. Yao,et al.  Fetal Leydig cells: progenitor cell maintenance and differentiation. , 2010, Journal of andrology.

[5]  R. Lovell-Badge,et al.  Somatic Sex Reprogramming of Adult Ovaries to Testes by FOXL2 Ablation , 2009, Cell.

[6]  Abdelkader Essafi,et al.  Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin , 2009, Nature Genetics.

[7]  Hao Chang,et al.  Overactive Beta-Catenin Signaling Causes Testicular Sertoli Cell Tumor Development in the Mouse1 , 2009, Biology of reproduction.

[8]  H. Scherthan,et al.  Testis cord differentiation after the sex determination stage is independent of Sox9 but fails in the combined absence of Sox9 and Sox8. , 2009, Developmental biology.

[9]  Hao Chang,et al.  Wt1 negatively regulates β-catenin signaling during testis development , 2008, Development.

[10]  N. Hastie,et al.  Wt1 and retinoic acid signaling are essential for stellate cell development and liver morphogenesis. , 2007, Developmental biology.

[11]  Ho-Joon Lee,et al.  Octylphenol reduces the expressions of steroidogenic enzymes and testosterone production in mouse testis , 2007, Environmental toxicology.

[12]  Daishu Han,et al.  The role of Tyro 3 subfamily receptors in the regulation of hemostasis and megakaryocytopoiesis. , 2007, Haematologica.

[13]  Fei Gao,et al.  The Wilms tumor gene, Wt1, is required for Sox9 expression and maintenance of tubular architecture in the developing testis , 2006, Proceedings of the National Academy of Sciences.

[14]  S. Nef,et al.  Prostaglandin D2 induces nuclear import of the sex‐determining factor SOX9 via its cAMP‐PKA phosphorylation , 2005, The EMBO journal.

[15]  N. Hastie,et al.  Development of an siRNA-based method for repressing specific genes in renal organ culture and its use to show that the Wt1 tumour suppressor is required for nephron differentiation. , 2003, Human molecular genetics.

[16]  C. Lécureuil,et al.  Sertoli and granulosa cell‐specific Cre recombinase activity in transgenic mice , 2002, Genesis.

[17]  K. Wagner,et al.  The Wilms' tumor gene Wt1 is required for normal development of the retina , 2002, The EMBO journal.

[18]  K. H. Albrecht,et al.  Evidence that Sry is expressed in pre-Sertoli cells and Sertoli and granulosa cells have a common precursor. , 2001, Developmental biology.

[19]  K. Naito,et al.  Characterization of mesonephric cells that migrate into the XY gonad during testis differentiation. , 2001, Experimental cell research.

[20]  Robin Lovell-Badge,et al.  Targeted Mutagenesis of the Endogenous Mouse Mis Gene Promoter In Vivo Definition of Genetic Pathways of Vertebrate Sexual Development , 1999, Cell.

[21]  C. Englert,et al.  The Wilms tumor suppressor gene wt1 is required for development of the spleen , 1999, Current Biology.

[22]  A. Schedl,et al.  YAC complementation shows a requirement for Wt1 in the development of epicardium, adrenal gland and throughout nephrogenesis. , 1999, Development.

[23]  B. Capel,et al.  Sertoli cells of the mouse testis originate from the coelomic epithelium. , 1998, Developmental biology.

[24]  N. Moreno-Mendoza,et al.  Mesonephric stromal cells differentiate into Leydig cells in the mouse fetal testis. , 1998, Experimental cell research.

[25]  A. McMahon,et al.  Sertoli cell signaling by Desert hedgehog regulates the male germline , 1996, Current Biology.

[26]  David Housman,et al.  WT-1 is required for early kidney development , 1993, Cell.

[27]  B. Williams,et al.  Inherited WT1 mutation in Denys-Drash syndrome. , 1992, Cancer research.

[28]  D. Housman,et al.  Germline mutations in the Wilms' tumor suppressor gene are associated with abnormal urogenital development in Denys-Drash syndrome , 1991, Cell.

[29]  D. Housman,et al.  Expression of the Wilms' tumor gene WT1 in the murine urogenital system. , 1991, Genes & development.

[30]  Peter Goodfellow,et al.  "Male Development of Chromosomally Female Mice Transgenic for Sry gene" (1991), by Peter Koopman, et al. , 2014 .

[31]  D. Housman,et al.  Evidence for WT1 as a Wilms tumor (WT) gene: intragenic germinal deletion in bilateral WT. , 1991, American journal of human genetics.

[32]  Peter Goodfellow,et al.  A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes , 1990, Nature.

[33]  A. Poustka,et al.  Homozygous deletion in Wilms tumours of a zinc-finger gene identified by chromosome jumping , 1990, Nature.

[34]  M. Griswold,et al.  DMRT1 prevents female reprogramming in the postnatal mammalian testis. , 2011, Nature.

[35]  S. G. Haider.  Cell biology of Leydig cells in the testis. , 2004, International review of cytology.

[36]  Philippe Soriano.  Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.